OBJECTIVE:To establish a method for the contents determination of emodin,rhein,aloe-emodin,chrysophanol, physcion,geniposide,chlorogenic acid in Yinchenhao decoction and its dispensing granule,and to compare the difference among the above-mentioned ingredients. METHODS:HPLC was adopted to determine the contents of emodin, rhein, aloe-emodin, chrysophanol and physcion:the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 254 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of geniposide by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-water(13∶87, V/V)at a flow rate of 1 ml/min,the detection wavelength was 238 nm,column temperature was 35 ℃,and injection volume was 20 μl. When determining the content of chlorogenic acid by HPLC,the column was Ecosil C18 with mobile phase of acetonitrile-0.5% phosphoric acid (10∶90,V/V) at a flow rate of 1 ml/min,the detection wavelength was 327 nm,column temperature was 35 ℃,and injection volume was 20 μl. RESULTS:The linear range was 2-20 μg/ml for emodin(r=0.996 5),5.2-52.0 μg/ml for rhein(r=0.998 5),2.6-26.0 μg/ml for aloe-emodin(r=0.999 9),1.0-10.4 μg/ml for chrysophanol(r=0.999 9),1.0-10.0 μg/ml for physcion(r=0.999 8),20-200 μg/ml for geniposide(r=0.999 9),20-200 μg/ml for chlorogenic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 91.1%-96.9%(RSD=2.0%,n=6)for emodin、93.9%-96.1%(RSD=0.8%,n=6)for rhein、90.9%-93.4%(RSD=1.2%,n=6)for aloe-emodin、88.5%-92.7%(RSD=1.8%,n=6) for chrysophanol、82.1% -87.9%(RSD=2.5% ,n=6)for physcion,100.4% -102.0%(RSD=0.7% ,n=6)for geniposide、101.1%-102.2%(RSD=0.4%,n=6) for chlorogenic acid. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous contents determination of active ingredients in Yinchenhao decoction and its dispensing granule. The contents of active ingredients in Yinchenhao dispensing granule are obviously higher than those in its decoction.

Objective To evaluate the clinical application of enteral nutrition by nasojejunal tube insertion and by percutaneous endoscopic gastrostomy (PEG) in elderly patients.Methods A total of 65 elderly patients with dysphagia recruited at our department from January 2010 to November 2014 were divided into the nasojejunal tube feeding group (35 cases) and the PEG feeding group (30 cases).Differences between these two groups in nutritional indexes,immunological indexes,complications and mortality were analyzed retrospectively.Results Serum total protein,albumin and prealbumin and upper arm circumferences all increased after treatment with nasojejunal tube feeding or percutaneous endoscopic gastrostomy (P＞0.05).There was overall improvement in nutritional status,as assessed by Nutritional Risk Screening 2002 (NRS2002).Specifically,the before/one month-after-treatment ratio of scores was 3.72±0.91/1.90±0.61 (t=7.24,P＜0.01) for the nasojejunal tube feeding group and 3.52±1.23/2.02±0.53 (t=4.17,P＜0.01) for the PEG feeding group.Compared with NRS2002 scores at one month post-operation,further improvement was achieved at 3 months postoperation both for the nasojejunal tube feeding group (1.89±0.65,t=5.21,P＜0.01) and for the PEG feeding group (1.91±0.62,t=4.40,P＜0.01).There was no difference in the indexes of nutrition,immune status or mortality between the two groups (P＞0.05).Although improvement in CD3+,CD4+,CD8+,CD4+/CD8+,IgA,IgG,and IgM was seen in both groups after operation,the differences did not reach statistical significance (P＞0.05).The incidence of aspiration pneumonia was notably lower (P＜0.05) while the incidence of diarrhea was much higher (P＜0.05) in the nasojejunal tube feeding group than in the PEG feeding group at one month and three months.The two groups had similar causes of death and mortality rates.Conclusion Both nasojejunal tube and PEG feeding can improve the nutritional status of elderly patients with dysphagia.However,the choice for the route of nutrition should be individualized.

Objective: To establish the method of percutaneous endoscopic gastrostomy(PEC) and percutaneous endoscopic jejunostomy (PEJ) for enteral nutrition. Methodes: PEG tubes were placed in 114 patients with Pull method. On the foundation of PEG, PEJ tubes were placed in 26 patients by pushing endoscopy to send tubes through Treitz ligment with usingthe the clip. Results: All PEG insertion was performed successfully. PEJ tubes were placed successfully with a new method in 26 patients. 15 patients had a little blooding and 8 patients had slight infection. 21 patients had respiratory tract infection and had been cured by using antibiotic. There was no severe complication. Conclusion: PEG is simple、safe、efficient. The new method of PEJ is feasible.

BACKGROUND:In recent years, the application of stem cel s to treat autoimmune diseases has become a hot spot. But, studies on umbilical cord blood mesenchymal stem cel s transplantation for the treatment of polymyositis/dermatomyositis are rarely reported. OBJECTIVE:To explore the immunologic mechanism of Th cytokines on the occurrence and development of polymyositis/dermatomyositis by observing the changes in serum interferon-γ, interleukin-4 and interleukin-17 in patients after umbilical cord blood mesenchymal stem cel s transplantation. METHODS:Eighty-one polymyositis/dermatomyositis patients were selected and divided into conventional therapy group (n=44) undergoing glucocorticoid and immunosuppressants therapy and cel transplantation group (n=37) undergoing intravenous infusion of umbilical cord blood mesenchymal stem cel s at a density of (3.5-5.2 )×107 . Dosing regimen was same in the two groups. After fol ow-up of 1, 3, 6 months, the changes of creatine kinase and myodynamia were evaluated;after fol ow-up of 3 and 6 months, lung imaging was evaluated;in the cel transplantation group, interferon-γ, interleukin-4 and interleukin-17 levels were detected before treatment and at 3 and 6 months after treatment. RESULTS AND CONCLUSION:At 1, 3, 6 months after treatment, the creatine kinase level was significantly decreased, and the muscle force grade was significantly increased in both groups (both P<0.001). Compared with the conventional therapy group, the creatine kinase level was lower and the muscle force grade was higher in the cel transplantation group (both P<0.001). Results from lung imaging test showed a remarkable improvement after cel transplantation, and it indicated that umbilical cord blood mesenchymal stem cel s transplantation had good stability. At 6 months after transplantation, the level of interferon-γwas significantly increased, while the interleukin-4 level was decreased significantly (both P<0.01);at 3, 6 months after cel transplantation, the levels of interleukin-17 were significantly decreased (P<0.01). Levels of interleukin-4 and interleukin-17 were positively correlated with the level of creatine kinase at 6 months after cel transplantation (r=0.467, 0.488, both P<0.05), but there was no obvious correlation between the levels of interferon-γand creatine kinase (r=0.213, P>0.05). These findings indicate umbilical cord blood mesenchymal stem cel s transplantation combined with glucocorticoid and immunosuppressants therapy can adjust immune network effects and improve the immune tolerance in polymyositis/dermatomyositis patients, which is safe and effective.

Objective To establish a new method based on Taqman's fluorescent ARMS-PCR technique for detecting the KRAS mutations in human paraffin embedding tissue samples,which is a reference value of clinical medication.Methods Specific primers and probes were designed based on the conserved sequences of KRAS and internal reference genes(GAPDH).Taqman's fluorescent ARMS-PCR detection systems and sequencing systems were established for KRAS detection.In this paper,paraffin embedded tissue samples of 200 patients with colorectal cancer(CRC)are detected and compared with gold standard Sanger sequencing.The results of KRAS gene mutation are statistically analyzed,and the consistency of the two detection methods is analyzed.Results This method is able to detect KRAS gene mutations in human paraffin embedding tissue samples.And the results are 100％consistent with Sanger sequencing.Conclusion The method based on Taqman's fluorescent ARMS-PCR established in this research can detect KRAS mutations simply,quickly and accurately,which is valuable for clinical medication and suitable for clinical application.

Objective The three-dimensional finite element model of the normal ankle joint was established to simulate the changes of stress and displacement under stress from different directions and of different magnitudes so as to provide a theoretical basis for the biomechanical mechanism of the ankle joint injury.Methods Spiral CT scan was performed on the left ankle of a 30 year old healthy male volunteer to obtain the original CT image data.The three-dimensional digital model of ankle joint generated by Mimics and Geomagic softwares was imported into the software Ansys.The three-dimensional finite element model of ankle joint with complete anatomical structure was obtained after the main steps of meshing,central node,element linking and module loading using finite element method.Stress from different directions and of different magnitudes were loaded unto the model.The stress changes were measured by foot stress distribution measurement system.The stress changes,displacement change distribution,the stress peak value of heel are,metatarsal stress,and plantar contact stress area as well as the maximum,minimum,and contact are of the tibial articular surface contact stress were compared between the finite element model and the volunteer himself to verify the consistency.Results For the finite element model of normal ankle joint,the plantar peak stress was [(0.33 ± 0.10) MPa],the metatarsal stress was [(0.13 ± 0.21)MPa],the foot contact stress area was [(78.60 ±0.32)mm2],the tibial articular surface maximum contact stress was [(2.72 ± 0.10) MPa],the minimum contact stress was [(1.35 ±0.12)MPa],and the contact stress area was [(79.1 ± 0.14)mm2].For the volunteer,double foot plantar peak stress was [(0.35 ± 0.12)MPa],the metatarsal stress [(0.13 ±0.16)MPa],the foot contact stress area was [(77.3 ± 0.42)mm2],the tibial articular surface maximum contact stress was [(2.79 ± 0.23) MPa],the minimum contact stress was [(1.37 ± 0.20) MPa],and the contact stress area was [(79.10 ±0.14)mm2] (P ＜0.05).Therefore,the three finite element model of ankle joint was basically consistent with the real human ankle joint because of the similiar distribution,trend,and values.Conclusion The three-dimensional finite element model of normal ankle joint can objectively reflect the anatomical structure and biomechanical characteristics of the joint,which is of great value to understand the changes of the internal mechanics and ankle joint injury.

From January 2016 to June 2017, 68 patients with thoracic osteoporotic compression fractures were treated with percutaneous kyphoplasty, including 31 cases with ultrasound-guided thoracic paravertebral nerve block (group A) and 37 cases with local anesthesia (group B). The duration of analgesia in group A was longer than that in group B (P<0.05). The satisfaction of anesthesia in group A was higher than that in group B (90% vs. 68%, P<0.05). There was no significant difference in length of hospital stay and cost between the two groups (P>0.05). The post-operative VAS scores was significantly lower than those of pre-operation in both groups (P<0.05). The intraoperative VAS score of group A was lower than that of group B (P<0.05). There was no significant difference between the two groups in mean arterial pressure and heart rate (P>0.05). No cardiovascular and cerebrovascular adverse reactions occurred in both groups. Ultrasound-guided thoracic paravertebral nerve block is a safe and effective method used in percutaneous kyphoplasty.

Objective:To analyze the differences and similarities of pre-treatment and post-treatment lung microbiome of acute respiratory distress syndrome (ARDS) and find out the change rules of the lung microbiome in the progression of ARDS according to different prognosis.Methods:A retrospective study was conducted. Patients with ARDS caused by severe pneumonia admitted to intensive care unit (ICU) of Jiangmen Central Hospital from February 2019 to January 2020 were enrolled as the study subjects. The patients were divided into pre-treatment (ARDS-preT) group (24 cases), post-treatment survival (ARDS-poT-Survival) group (17 cases), and post-treatment death (ARDS-poT-Dead) group (7 cases). ICU patients with mild pulmonary infection and non-ARDS admitted to ICU during the same period were enrolled as control group (25 cases). The similarities and differences of lung microbiome in four groups were analyzed and compared, and the possible pathogenic bacteria (potential risk factors for death) and probiotics (potential survival and protective factors) related to death caused by ARDS were screened.Results:In terms of pathogenic microorganisms, the positive rates of Escherichia coli and Candida albicans in the ARDS-poT-Dead group were significantly higher than those in the ARDS-poT-Survival group [57.1% (4/7) vs. 5.9% (1/17) and 57.1% (4/7) vs. 0% (0/7), both P < 0.05]. In the screening of background bacteria, the decrease of bacteria in the ARDS-preT group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the control group, the reduced bacteria might be pulmonary probiotics (potential protective factor for ARDS). The screening result was Hydrobacter [ARDS-preT group vs. ARDS-poT-Survival group: 62.5% (15/24) vs. 94.1% (16/17); ARDS-poT-Dead group vs. ARDS-poT-Survival group: 14.3% (1/7) vs. 94.1% (16/17); ARDS-poT-Dead vs. control: 14.3% (1/7) vs. 96.0% (24/25), all P < 0.05]. In the screening of background bacteria, the increase of bacteria in the ARDS-poT-Dead group compared with the ARDS-preT group, the ARDS-poT-Dead group compared with the ARDS-poT-Survival group, the ARDS-poT-Dead group compared with the control group, and the increased bacteria might be potential pulmonary pathogen (potential risk factor for death of ARDS), which belonged to Enterobacteria: Edwardsiella, Enterobacteriaceae, Escherichia, Klebsiella, Kluyvera, Lelliottia, Pantoea, Raoultella. Conclusions:The results revealed the increase of Escherichia coli or Candida albicans in pulmonary pathogenic microorganisms, or the increase of Enterobacteria in background bacteria may be the risk factors for the death of ARDS. Additionally, background bacteria Hydrobacter probably is a protective factor for the survival of ARDS. Whether it can be used as a novel treatment for ARDS is worth further investigation.

Objective:To investigate the effects of insulin resistance induced by long-term high-fat diet on cognitive function and brain phosphorylated Tau protein in pR5 MAPT Tau transgenic mice.Methods:Eight-week-old female pR5 MAPT transgenic mice were divided into standard diet(STD) group (pR5 STD, n=8) and high-fat diet(HFD) group (pR5 HFD, n=8). Female wild-type C57BL/6 mice fed with STD were used as control group (WT STD, n=8). The experiment was carried out for 30 weeks until the mice were old.During the experiment, the weight of mice was measured once a week and fasting blood glucose was measured once every two weeks.Thirty weeks later, glucose tolerance test and insulin tolerance test were carried out.Forced swimming test and tail suspension test were used to evaluate the depressive behavior of mice, and elevated plus maze test was used to evaluate the anxiety behavior, Morris water maze test was used to evaluate spatial memory behavior.The levels of total Tau protein and phosphorylated Tau proteins H7-tau, p-tau-Ser396 and p-tau-Thr231 were detected by Western blot.SPSS 17.0 software was used for statistical analysis, repeated measurement ANOVA was used for the data of glucose tolerance test and insulin tolerance test, one-way ANOVA was used for multi group comparison, and LSD test was used for further pairwise comparison. Results:After 30 weeks of high-fat diet, there were significant differences in body weight and fasting blood glucose among the three groups ( F=808.31, 1 117.18, both P<0.01). The body weight of mice in pR5 HFD group ((54.35±2.52)g) was higher than those in pR5 STD group ((24.95±1.15) g) and WT STD group ((23.86±1.10) g) (both P<0.01), and the fasting blood glucose of mice in pR5 HFD group ((8.12±0.24) mmol/L) was significantly higher than those in pR5 STD group ((4.64±0.13) mmol/L) and WT STD group ((4.45±0.22) mmol/L) (both P<0.01). Glucose tolerance test showed that within 120 minutes after injection of glucose, there was a significant time and group interaction in the blood glucose value among the three groups ( F=113.30, P<0.01). After glucose injection, the peak value of blood glucose in pR5 HFD group was delayed, suggesting that glucose tolerance in pR5 HFD group was impaired.The insulin tolerance test showed that there was a significant interaction between time and group in the insulin tolerance among the three groups ( F=209.92, P<0.01). After injection of insulin, the blood glucose in pR5 HFD group decreased slowly, reaching the valley value at 60 min, and then the blood glucose increased significantly, suggesting that the sensitivity of pR5 HFD group mice to insulin decreased significantly.There were significant differences in the percentage of forced swimming immobility time and tail suspension immobility time among the three groups ( F=37.05, 59.29, both P<0.01). The two indexes of pR5 STD group and pR5 HFD group were both higher than those of WT STD group (all P<0.01), and those of pR5 HFD group were both higher than those of pR5 STD group (both P<0.01). The results of elevated plus maze showed that there were significant differences in the activity distance and time in open arm among the three groups ( F=7.82, 10.37, both P<0.05) .The activity distance ((0.40±0.21) m) and activity time ((27.38±8.80) s) of pR5 HFD group were significantly lower than those of pR5 STD group ((2.31±1.74) m, (63.56±27.52)s) (both P<0.05). The space exploration test showed that the residence time in the target quadrant of pR5 HFD group ((15.56±1.16)s) was less than that of pR5 STD group((19.18±0.64)s)( P<0.01), and the time of entering the platform area of pR5 HFD group((1.43±0.06)s) was less than that of pR5 STD group((1.66±0.12)s)( P<0.01). Western blot showed that there were significant differences in the levels of total Tau protein, H7-tau, p-tau-Ser396 and p-tau-Thr231 protein among the three groups ( F=101.50, 80.60, 55.47, 30.89, all P<0.05). Further pairwise comparison showed that the levels of the four Tau proteins showed: the levels of the proteins in pR5 STD group and pR5 HFD group were all higher than those of WT STD group (all P<0.01), and those in pR5 HFD group were all higher than those in pR5 STD group (all P<0.05). Conclusion:Long-term high fat diet causes obesity, hyperglycemia and peripheral insulin resistance, and promotes the cognitive impairment of MAPT mice, which is related to the increase of Tau protein phosphorylation in the brains of MAPT mice.

The two forms of brain-derived neurotrophic factor (BDNF), mature BDNF (mBDNF) and BDNF precursor (proBDNF), bind to corresponding receptors and mediate a series of biological signals, which play important roles in proliferation, survival and apoptosis. More and more studies have focused on the role of unbalanced transformation from proBDNF to mBDNF in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, as well as their potential as biomarkers and treatment targets. This article reviews the mechanism and therapeutic application of BDNF and proBDNF/mBDNF imbalance in the above-mentioned neurodegenerative diseases to provide new perspective and approach for the pathogenesis, early diagnosis and treatment of neurodegenerative diseases such as AD and PD.